Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry
- Oregon Health and Science University, United States
- Creighton University, United States
- National Institute on Drug Abuse/National Institute on Alcohol Abuse and Alcoholism, United States
Abstract
Identifying neurons that have functional opioid receptors is fundamental for the understanding of the cellular, synaptic and systems actions of opioids. Current techniques are limited to post hoc analyses of fixed tissues. Here we developed a fluorescent probe, naltrexamine-acylimidazole (NAI), to label opioid receptors based on a chemical approach termed 'traceless affinity labeling'. In this approach, a high affinity antagonist naltrexamine is used as the guide molecule for a transferring reaction of acylimidazole at the receptor. This reaction generates a fluorescent dye covalently linked to the receptor while naltrexamine is liberated and leaves the binding site. The labeling induced by this reagent allowed visualization of opioid-sensitive neurons in rat and mouse brains without loss of function of the fluorescently labeled receptors. The ability to locate endogenous receptors in living tissues will aid considerably in establishing the distribution and physiological role of opioid receptors in the CNS of wild type animals.
Data availability
All data generated or analysed during this study are includes in the manuscript and supporting files.
Article and author information
Author details
Seksiri Arttamangkul
Funding
National Institute on Drug Abuse (DA008163)
- John T Williams
National Institute on Drug Abuse (DA048136)
- John T Williams
National Institute on Drug Abuse (DA042779)
- William Birdsong
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All animal uses were conducted in accordance with the National Institutes of Health guidelines and with approval from the Institutional Animal Care and Use Committee (IACUC) protocol #IP00000160 of the Oregon Health & Science University. Rats and mice were anesthetized with isofluorane before euthanized with minimal suffering.
Copyright
This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
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Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry
eLife 8:e49319.
https://doi.org/10.7554/eLife.49319
